Generation of NelfbdTAG mouse model to study the dTAG system in vivo.
(A) Schematic illustration of the proteasomal degradation in the dTAG system. The numbers represent the sequence of events from recognition of tag to degradation.
(B) Immunofluorescence of targeted mESCs showing nuclear localized HA signal corresponding to NELFB +/− 500nM dTAG-13 treatment for 30 mins. Nuclei labeled with Hoechst. Scale bar, 70μm.
(C) Western blot of targeted mES cells’ whole cell lysates +/− dTAG-13 treatment for indicated time intervals. Anti-NELFB antibody was used. 20ug of protein loaded/lane.
(D) Immunofluorescence of a single E7.5 litter from a heterozygous NelfbdTAG/+ male and wild-type Nelfb+/+ female showing germline transmission of the targeted allele. Nuclei labeled with Hoechst. Scale bar, 500μm.